Effects of commercial anthocyanin-rich extracts on colonic cancer and nontumorigenic colonic cell growth

Commercially prepared grape (Vitis vinifera), bilberry (Vaccinium myrtillus L.), and chokeberry (Aronia meloncarpa E.) anthocyanin-rich extracts (AREs) were investigated for their potential chemopreventive activity against colon cancer. The growth of colon-cancer-derived HT-29 and nontumorigenic colonic NCM460 cells exposed to semipurified AREs (10-75 microg of monomeric anthocyanin/mL) was monitored for up to 72 h using a sulforhodamine B assay. All extracts inhibited the growth of HT-29 cells, with chokeberry ARE being the most potent inhibitor. HT-29 cell growth was inhibited approximately 50% after 48 h of exposure to 25 microg/mL chokeberry ARE. Most importantly, the growth of NCM460 cells was not inhibited at lower concentrations of all three AREs, illustrating greater growth inhibition of colon cancer, as compared to nontumorigenic colon cells. Extracts were semipurified and characterized by high-pressure liquid chromatography, spectrophotometry, and colorimetry. Grape anthocyanins were the glucosylated derivatives of five different anthocyanidin molecules, with or without p-coumaric acid acylation. Bilberry contained five different anthocyanidins glycosylated with galactose, glucose, and arabinose. Chokeberry anthocyanins were cyanidin derivatives, monoglycosylated mostly with galactose and arabinose. The varying compositions and degrees of growth inhibition suggest that the anthocyanin chemical structure may play an important role in the growth inhibitory activity of commercially available AREs.     

Inhalant anesthetic requirement in cats is animal-dependent

Minimum alveolar concentration (MAC) of an inhalant is an indicator of its anesthetic potency. Individuals vary in their sensitivity to anesthetic agents as demonstrated by different individual MAC values. We hypothesized that individual animal sensitivity would be maintained with different inhalant anesthetics. As part of separate studies, six female DSH cats, aged 24 ± 2.5 (mean ± SD) months and weighing 3.5 ± 0.3 kg, were studied similarly on three separate occasions over a 12‐month period to determine the MAC of isoflurane (ISO), sevoflurane (SEVO), and desflurane (DES), respectively. In each study, chamber induction was followed by orotracheal intubation, and anesthesia was maintained via a nonrebreathing circuit. ECG, pulse oximetry, Doppler systolic blood pressure, end‐tidal gases, and esophageal temperature were monitored. End‐tidal gases were hand‐sampled from a catheter whose tip lay level with the distal end of the ET tube. Gases were analyzed by Raman spectrometry and, for each agent, the analyzer was calibrated with at least three gas standards. MAC was determined in triplicate using standard tail‐clamp technique. Data were analyzed by two‐way anova followed by Tukey's test and significant differences were found. Average MACs (%) for ISO, SEVO, and DES were 1.90 ± 0.18, 3.41 ± 0.65, and 10.27 ± 1.06, respectively. Body temperatures, Doppler systolic blood pressure, and SpO₂ were recorded at the time of MAC determinations for ISO, SEVO, and DES were 38.3 ± 0.3, 38.6 ± 0.1, 38.3 ± 0.35 °C; 71 ± 8, 75 ± 16, 88 ± 12 mm Hg; 99 ± 1, 99 ± 1, 99 ± 1%, respectively. Both the anesthetic agent and the individual cat had significant effects on MAC (p = 0.0001 and 0.0185, respectively). MAC varied between individuals and cats were consistent in their order of sensitivity to inhalant anesthetics across the three agents. Within this group of cats, the relationship of individual MAC to the group MAC for each of the three inhalant agents was maintained. This suggests that any individual may be consistently more or less sensitive to a variety of inhalant agents.     

NITROGEN,PHOSPHORUS, AND POTASSIUM EFFECTS ON GRAIN SORGHUM PRODUCTION AND STALK ROT FOLLOWING ALFALFA AND BIRDSFOOT TREFOIL

Grain sorghum [Sorghum bicolor (L.)], grown on the often infertile claypan soils of the eastern Great Plains, requires attention to soil fertility. Experimental objectives were to determine the effects of phosphorus (P) and potassium (K) fertility levels, N application, and legume residual on grain sorghum production and stalk rot. Following alfalfa and birdsfoot trefoil, first-year sorghum yield was 7 Mg ha^?1 and not affected by N fertilizer. In subsequent years, yield increases due to N were less than 20%. Sorghum yield increased at low P and K rates, especially with nitrogen (N) fertilization and was greater following birdsfoot trefoil than following alfalfa. In 1995 when fertilized with N, lodging and stalk rot severity were increased by P and reduced by K. In 1996, stalk rot severity was reduced by K fertilization. Grain sorghum, grown after legume crops, required minimal levels of P and K, especially when N fertilizer was added.     

Calibration of two techniques for estimating herbage mass

Two non-destructive techniques for estimating herbage mass (dry matter yield) were evaluated during the growing season of 1988, from April to September. Samples were collected from four sites, with ten observations made on twenty-five days at each site, giving a total of 612 sample units. Observations were obtained from each unit, either with a plastic rising-plate meter (335) or with a metal rising-plate meter (227). In addition, measurements with a single-probe capacitance meter (Pasture Probe) were also made on all units. The usefulness of the non-destructive methods for predicting the actual yield was evaluated by calibrating them by means of regression on the yield determined by mowing 1·5-m² quadrats, weighing, subsampling and drying the grass. The yield measurements were logarithmically transformed to correct for the proportionality of the residual variance with the mowed dry matter yield. Logarithmic transformation of the sensor measurements improved the linearity of the calibration relationships. The combination of one rising-plate meter and the Pasture Probe resulted in a linear and additive model with a larger adjusted R² than models with just one of these two sensors. The addition of the predictors ‘external moisture’ (e.g. dew), ‘day number after mowing’ and ‘site number’ significantly increased the goodness of fit of many models. In some models addition of the logarithm of the ‘dry matter percentage’ was significant. Other predictors in these models such as “Julian day number” and “grazing” proved not to be significant. The adjusted R²s in the best models for the metal and the plastic discs were 89·7% and 87·3% respectively. The corresponding coefficients of variation were 26·2% and 26·1%. The results show that further research is necessary to refine the techniques or to develop new techniques to estimate the dry matter yield, before the sensors can be usefully integrated into a management system.     

Implementation of simple sequence repeat markers to genotype Florida strawberry varieties

Simple sequence repeats (SSRs or microsatellites) are frequently used as a robust, rapid, and relatively inexpensive means of genotyping. Recent reports in cultivated strawberry (Fragaria × ananassa) have identified a widely applicable set of SSRs that permits discrimination of closely-related genotypes. In the present study this same set of SSRs is analyzed in cultivars commonly grown in the state of Florida, as well as advanced breeding selections from the University of Florida program. Nine primer pairs have been used to produce discrete SSR patterns for all lines tested, including diagnostic sets that distinguish between closely-related cultivars and/or breeding selections. A comparison of markers to known pedigrees is presented, along with analysis of relative relatedness between lines studied. We also detail important technical considerations for transferability and limitations of these technologies between laboratories. The resulting genotype data for cultivars commonly used in Florida are accessible as supplemental data in graphic format and allow comparison with other cultivars. Users can thus select the most diagnostic SSRs to test the lines in question. These resources provide new tools for breeders and nurseries to authenticate genotypes, follow inheritance patterns and enforce patent protection. As important, this report underscores the strengths and weaknesses of applying the original methodologies to a different plant population in independent laboratories.     

White-tailed deer (Odocoileus virginianus) develop spirochetemia following experimental infection with Borrelia lonestari

Borrelia lonestari is considered a putative agent of southern tick-associated rash illness (STARI) and is known to occur naturally only in lone star ticks (Amblyomma americanum) and white-tailed deer (Odocoileus virginianus). We used a low passage isolate of B. lonestari (LS-1) to inoculate white-tailed deer, C3H mice, Holstein cattle, and beagles. Animals were monitored via examination of Giemsa and acridine orange stained blood smears, polymerase chain reaction (PCR), indirect fluorescent antibody (IFA) test, and/or culture isolation. Spirochetes were visualized in blood smears of both deer on days post-inoculation (DPI) 6, 8, 12 and one deer on DPI 15. Whole blood collected from deer tested PCR positive starting on DPI 4 and remained positive as long as DPI 28. Both deer developed antibody titers of >64, with a maximum IFA titer of 1024. The organism was reisolated from the blood of both deer on DPI 6 and one deer on DPI 12. All isolation attempts from mice, calves, or dogs were negative, although one of seven mice was transiently PCR positive. Mice and dogs developed an IFA titer > or =64, while calves lacked a detectable antibody response. These preliminary experimental infection trials show that white-tailed deer are susceptible to infection with B. lonestari and develop a spirochetemia following needle-inoculation, while C3H mice, calves, and dogs do not. Results suggest that deer may serve as a vertebrate reservoir host. Tick transmission studies are needed to confirm that this organism can be maintained in a natural cycle involving deer and A. americanum.